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Concentrating Protein with centrifuge concentrator but 50% glycerol!!?
Started by bc87, Oct 01 2009 06:21 AM
2 replies to this topic
#1
bc87
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Posted 01 October 2009 - 06:21 AM
I need to increase the concentration of my protein by a 1/2 but problem is - it's in a buffer with 50% glycerol. I'm using the Amicon centrifuge devices and I knew it was going to be slow. After the first 25 min spin I got a sad <1mL in the flowthrough but after the 2nd 25min spin...doesn't look like a drop went through. Is this actually a feasible way to concentrate this protein?......
#2
mdfenko
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Posted 02 October 2009 - 10:12 AM
it should work okay, glycerol will pass through the membrane.
you may need to clear the membrane (flush it with the sample), it may have "clogged".
you may need to clear the membrane (flush it with the sample), it may have "clogged".
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genius does what it must
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#3
Prep!
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Posted 26 October 2009 - 08:22 PM
mdfenko, on Oct 2 2009, 11:42 PM, said:
it should work okay, glycerol will pass through the membrane.
you may need to clear the membrane (flush it with the sample), it may have "clogged".
you may need to clear the membrane (flush it with the sample), it may have "clogged".
check the rpm that you are using... and after each spin pipette the remaining solution and mix it so that any clogged surface near the membrane is washed too!!! i presume you are using 15 ml amicons with double filters... if not use that cause in that case it should be faster!!
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